Differentiation inducer to brown-like adipocyte of white adipocyte
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example 1
Generation and Isolation and Refining of UHA1028
[0082]1 g of trans-resveratrol and 1 g of sinapic acid (manufactured by Wako Pure Chemical Industries, Ltd.) were dissolved in 20 mL of ethanol, and then 20 mL of mineral water was added, thereby obtaining a solution containing the resveratrol and the sinapic acid (pH=4.9). The solution containing the resveratrol and the sinapic acid was heated at 130° C. for 90 minutes in an autoclave (“SANYO LABO AUTOCLAVE”, manufactured by SANYO Electric Co., Ltd.). 1 mL of the obtained reaction solution was taken out, diluted with methanol to 50 mL in a measuring flask, and then 10 μL of the resultant solution was analyzed by HPLC.
[0083]The HPLC analysis was carried out under the following conditions.
Column: Reverse phase column “Develosil (Registered Trademark)
C-30-UG-5” (4.6 mmi.d.×250 mm)
[0084]Mobile phase: A . . . H2O (0.1% trifluoroacetic acid (TFA)),[0085]B . . . Acetonitrile (0.1% TFA)
Flow velocity: 1 mL / min
Injection: 10 μL
Detection: 254 nm
[...
example 2
Verification of Expression of Indicator Gene of Differentiation Induction Action to Brown-Like Adipocytes of White Adipocytes of UHA1028
[0092]In order to evaluate the differentiation induction action to brown-like adipocytes of white adipocytes, the evaluation was carried out using 3T3-L1 cells (mouse-derived preadipocytes). The 3T3-L1 preadipocytes are usually differentiated to white adipocytes through a differentiation induction process to be matured. However, by the differentiation induction to brown-like adipocytes, an expression of Cidea gene, an increase in the expression amount of Cox7a1 gene expressed in mitochondria, an increase in the expression amount of cytochrome c oxidase protein specifically expressed in mitochondria, and an expression enhancement of UCP1 gene, which are hardly observed in white adipocytes, are observed. Then, the differentiation induction to brown-like adipocytes of white adipocytes was confirmed based on the expression amount of each gene of Cidea, ...
example 3
Verification of Expression of Indicator Protein of Differentiation Induction Action to Brown-Like Adipocytes of White Adipocytes of UHA1028
[0102]In order to evaluate the differentiation induction action to brown-like adipocytes of white adipocytes, the evaluation was carried out using 3T3-L1 cells (mouse-derived preadipocytes). The 3T3-L1 preadipocytes are usually differentiated to white adipocytes through a differentiation induction process to be matured. However, by the differentiation induction to brown-like adipocytes, an increase in the expression amount of cytochrome c oxidase protein specifically expressed in mitochondria is observed. Then, the differentiation induction to brown-like adipocytes of white adipocytes was confirmed based on the expression amount of the CO1 protein.
[0103]Culture of cells was carried out by the same method as that of Example 2 using the same samples and cells as those in Example 2. After the end of the culture, 400 μL of RIPA buffer (manufactured b...
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